title: Expression, purification and crystallization of the FP domain of the human F-box protein Fbxo7.
abstract: Fbxo7 is a conserved protein in higher eukaryotes that belongs to the F-box protein family. Fbxo7 is the substrate-recognition component of the SCFFbxo7 (Skp1-Cul1-Fbox protein) E3 ubiquitin ligase. Besides the F-box motif, Fbxo7 also contains a C-terminal proline-rich region, an N-terminal ubiquitin-like domain and a novel FP (Fbxo7/PI31) domain preceding the F-box motif. The FP domains of Fbxo7 and the PI31 proteasome inhibitor mediate interaction between the two proteins. For structure determination of the FP domain of Fxbo7, a protein construct (amino acids 181-335) corresponding to the FP domain was expressed, purified and crystallized. The native and selenomethionine-labeled proteins crystallized in different crystal forms. Native and single-wavelength anomalous dispersion data sets with diffraction to 2.1 and 2.0 Å resolution, respectively, have been collected and structure determination is in progress.
authors.affiliation: Chemistry and Biochemistry, Southern Illinois University, 1245 Lincoln Drive, Carbondale, IL 62901, USA.
authors.forename: Jinsai
authors.initials: J
authors.lastname: Shang
authors.forename: Guan
authors.initials: G
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Xiaolan
authors.initials: X
authors.lastname: Huang
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Du
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D044783:F-Box Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D006801:Humans; D017434:Protein Structure, Tertiary
chemical_list: D044783:F-Box Proteins; C489149:FBXO7 protein, human
keywords: F-box; Fbxo7; Fbxo7/PI31 domain; PI31 proteasome inhibitor; SCF E3 ubiquitin ligase